Department of Pathology and Laboratory Medicine, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
Department of Microbiology and Infectious Disease, Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2023;2621:325-337. doi: 10.1007/978-1-0716-2950-5_19.
Submicroscopic malaria diagnosis requires highly sensitive tools instead of the conventional microscopy and rapid diagnostic tests (RDTs). While polymerase chain reaction (PCR) is more sensitive than RDTs and microscopy, the required capital cost and technical expertise hinder implementation of PCR in low- and middle-income countries. This chapter describes an ultrasensitive reverse transcriptase loop-mediated isothermal amplification (US-LAMP) test for malaria with a high sensitivity and specificity, while also being practical to implement in low-complexity laboratory settings. The workflow combines a silica spin column-based total nucleic extraction from dried blood spots (DBS) with US-LAMP amplifying the Plasmodium (Pan-LAMP) target and subsequent identification Plasmodium falciparum (Pf-LAMP).
亚显微疟疾诊断需要高灵敏度的工具，而不是传统的显微镜和快速诊断检测（RDT）。虽然聚合酶链反应（PCR）比 RDT 和显微镜更敏感，但所需的资本成本和技术专业知识阻碍了 PCR 在中低收入国家的实施。本章描述了一种超灵敏逆转录环介导等温扩增（US-LAMP）检测疟疾的方法，该方法具有高灵敏度和特异性，同时也适用于在低复杂性实验室环境中实施。该工作流程结合了基于二氧化硅的离心柱从干血斑（DBS）中提取总核酸，与 US-LAMP 扩增疟原虫（Pan-LAMP）靶标，随后鉴定恶性疟原虫（Pf-LAMP）。
